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Screening for benfluorex and its major urinary metabolites in routine doping controls

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Abstract

Benfluorex [1-(m-trifluoromethylphenyl)-2-(β-benzoyloxyethyl)aminopropane] has been widely used for the treatment of atherogenic metabolic disorders and impaired carbohydrate metabolism (particularly in obese type-II diabetic patients) as well as an anorectic drug. Due to its potentially performance-enhancing properties, benfluorex has been added to the list of prohibited compounds and methods of doping by the World Anti-Doping Agency (WADA) in 2010, necessitating the implementation of the drug as well as its major metabolites into routine doping control procedures. In the present study, human urinary metabolites of benfluorex were characterized by gas chromatography–electron ionization–mass spectrometry (GC-EI-MS) as well as liquid chromatography–electrospray ionization–high resolution/high accuracy tandem mass spectrometry (LC-ESI-MS/MS). Commonly employed sports drug testing approaches consisting of liquid–liquid extraction followed by GC-MS or urine dilution and immediate LC-MS/MS analysis were expanded and validated with regard to specificity, recovery (48–54%, GC-MS only), intra- and interday precision (<25%), limits of detection (5–8 ng/mL for LC-MS/MS and 80 ng/mL for GC-MS), and ion suppression (for LC-ESI-MS/MS only) to allow the detection of benfluorex metabolites 1-(m-trifluoromethylphenyl)-2-(2-hydroxyethyl)aminopropane (M1), 1-(m-trifluoromethylphenyl)-2-(2-carboxymethyl)aminopropane (M2), and 1-(m-trifluoromethylphenyl)-2-aminopropane (M3) as well as the glucuronic acid conjugate of M1.

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Acknowledgments

The study was conducted with support of the Federal Ministry of the Interior of the Federal Republic of Germany, the Ministry of Sport and Tourism of the Republic of Poland, and the Manfred-Donike Institute for Doping Analysis (Cologne, Germany).

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Authors and Affiliations

  1. Institute of Biochemistry—Center for Preventive Doping Research, German Sport University Cologne, Am Sportpark Müngersdorf 6, 50933, Cologne, Germany

    Mario Thevis, Gerd Sigmund, Vassilios Gougoulidis, Simon Beuck, Andreas Thomas & Wilhelm Schänzer

  2. Institute of Organic Chemistry, University of Cologne, Greinstraße 4, 50939, Cologne, Germany

    Nils Schlörer

  3. Department of Anti-Doping Research, Institute of Sport, Trylogii 2/16, 01-982, Warsaw, Poland

    Dorota Kwiatkowska & Andrzej Pokrywka

Authors
  1. Mario Thevis
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  2. Gerd Sigmund
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  3. Vassilios Gougoulidis
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  4. Simon Beuck
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  5. Nils Schlörer
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  6. Andreas Thomas
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  7. Dorota Kwiatkowska
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  8. Andrzej Pokrywka
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  9. Wilhelm Schänzer
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Correspondence to Mario Thevis.

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Published in the special issue Anti-Doping Analysis with Guest Editor Mario Thevis

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Thevis, M., Sigmund, G., Gougoulidis, V. et al. Screening for benfluorex and its major urinary metabolites in routine doping controls. Anal Bioanal Chem 401, 543–551 (2011). https://doi.org/10.1007/s00216-010-4455-4

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  • DOI: https://doi.org/10.1007/s00216-010-4455-4

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